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Updated: May 2, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
A comparative analysis of skin bacteria in healthy individuals through traditional cultivation and high-throughput
Jing Feng1, Shaolu Zhou1, Jiancong Huang1
1Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, State Key Laboratory of Applied Microbiology Southern China, Guangdong Detection Center of Microbiology, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, 510070, China.
Abstract:
The human skin microbiota is intricately linked to the microenvironmental niche of its habitat. Different skin sites may harbor distinct bacterial community structures due to their unique microenvironments. To elucidate the site-specific distribution patterns of skin bacterial communities, this study employed an integrated approach combining traditional microbial cultivation methods with 16S rRNA gene high-throughput sequencing to analyze 105 skin swab samples collected from the forehead, volar forearm, and scalp of 35 healthy individuals. A total of 675 bacterial strains encompassing 41 species and 18 genera were isolated by traditional culture-based methods. Staphylococcus and Cutibacterium were the dominant genera in different skin sites, with S. capitis, S. epidermidis, S. hominis, and C. acnes identified as the predominant species. High-throughput sequencing analyses revealed that the forehead and forearm harbored similar microbiota profiles, while the scalp displayed a more distinct bacterial community characterized by reduced alpha diversity and elevated relative abundances of Cutibacterium, Staphylococcus, and Lawsonella genera. The findings from both traditional cultivation methods and high-throughput sequencing suggested that the human skin bacterial community exhibits strong site-specificity. In contrast to the significant variations observed across different skin sites, gender and age exerted minimal effects on microbiota composition. These results suggest that microenvironmental factors play a more substantial role than age- and gender-related factors in shaping the diversity and community structure of the human skin microbiota.
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